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Non-destructive monitoring of root biomass in hydroponically grown leafy vegetables: comparison between machine learning-based RGB and hyperspectral imaging.

Plant Methods · 10 Mar 2026 · 10.1186/s13007-026-01515-8

Abstract

BACKGROUND: Root biomass serves as a critical indicator of plant eco-physiological status and crop productivity, yet its non-destructive monitoring remains challenging because of its underground location. The use of transparent nutrient film technique (NFT) systems enables direct observation of entire root systems, rendering image-based phenotyping feasible. In this study, we investigated and compared the performance of RGB and hyperspectral imaging for predicting root dry weight in hydroponically grown spinach (Spinacia oleracea L.). RESULTS: Using 430 root segments divided from 60 plants, three models were developed: (1) an area-based regression based on root coverage, (2) a convolutional neural network (CNN) using RGB images, and (3) a partial least squares regression (PLSR) model using hyperspectral data (450-950 nm). The area-based regression exhibited limited accuracy (R² = 0.446) because of saturation at high root coverage. The CNN model improved predictive performance (R² = 0.739) but tended to overestimate sparse roots as a result of resolution constraints. The PLSR model achieved the highest accuracy (R² = 0.822, RMSE = 0.019 g/segment), with significantly lower error than RGB-based approaches (P < 0.01). Variable importance in projection analysis indicated that PLSR effectively exploited spectral signatures at 450 nm (background contrast) and 750 nm (tissue scattering), thereby maintaining stable accuracy across the full biomass range. When validated using 104 independent plants, the PLSR model achieved high predictive accuracy. Furthermore, as a proof of concept, this model successfully visualized the spatiotemporal dynamics of root biomass accumulation over 50 days, with only a 7.70% relative error at harvest. CONCLUSIONS: To our knowledge, this study is among the first to demonstrate the non-destructive monitoring of biomass distribution within entire root systems under production conditions. Hyperspectral imaging combined with PLSR outperforms RGB-based approaches by capturing spectral signatures that reflect internal tissue properties of roots, thereby overcoming limitations caused by morphological occlusion. This approach provides a robust tool for precision agriculture and high-throughput phenotyping, enabling continuous assessment of root growth through simple modifications to the existing hydroponic systems.

Plant phenotyping relevance

RGB・ハイパースペクトル画像と機械学習/PLSRを用いて根乾物重を非破壊推定・検証する方法研究であり、植物表現型の取得と定量化が中心である。

abstractThe use of transparent nutrient film technique (NFT) systems enables direct observation of entire root systems, rendering image-based phenotyping feasible.
abstractIn this study, we investigated and compared the performance of RGB and hyperspectral imaging for predicting root dry weight in hydroponically grown spinach (Spinacia oleracea L.).
abstractWhen validated using 104 independent plants, the PLSR model achieved high predictive accuracy.
abstractThis approach provides a robust tool for precision agriculture and high-throughput phenotyping, enabling continuous assessment of root growth through simple modifications to the existing hydroponic systems.

Code and data availability

The paper's Data availability statement deposits the paper-specific phenotyping assets (raw hyperspectral images, RGB images, and root dry weight measurements) in a Zenodo record. The provided URL includes a token and 'preview=1', suggesting the record may not yet be fully open, but it is the authors' stated public URL

Datasetpublic

The datasets generated and analyzed during the model construction of the current study are available in the Zenodo repository: [https://zenodo.org/records/18072801?preview=1&token=eyJhbGciOiJIUzUxMiJ9.eyJpZCI6ImFiYTkzMzY2LTIzZjktNDlkMy1iZTBjLTk3M2E5YTUyOTFmZCIsImRhdGEiOnt9LCJyYW5kb20iOiIzNGE1ZjMxNDZhYjhiYjlhZWRiOWFjNzBkNzcwY2I3NyJ9.uR4HfosoSaVWhtSblMOS1v9bJFA5MvHwXvcW9uoNbcTWRDU4RNxZpVHjXTC3ulBM1JTlBbeHp_4T5EcILawxdg].The dataset includes: - Raw hyperspectral images and data- RGB images - Root dry weight measurements

Open resource ↗Zenodo · 18072801 · lines:176-248

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